中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
8期
1295-1298
,共4页
王永刚%陆满清%李佩瑜%颜志平%刘凌晓%王建华
王永剛%陸滿清%李珮瑜%顏誌平%劉凌曉%王建華
왕영강%륙만청%리패유%안지평%류릉효%왕건화
肝肠钙粘连蛋白%单克隆抗体%癌,肝细胞
肝腸鈣粘連蛋白%單剋隆抗體%癌,肝細胞
간장개점련단백%단극륭항체%암,간세포
Liver-intestine cadherin%Monoclonal antibody%Carcinoma,hepatocellular
目的 观察抗肝肠钙粘连蛋白(CDH17)单克隆抗体Lic5对肝癌细胞生物学行为的影响.方法 Western blot和实时定量聚合酶链反应(PCR)检测细胞株MHCC97H、MHCC97L、PLC/PRF/5及MIHA中CDH17的表达.噻唑蓝(MTT)比色法、细胞划痕法、Transwell法及平板克隆法检测Lic5对肝癌细胞生物学行为的影响.结果 CDH17仅在细胞株MHCC97H、MHCC97L中表达,Lic5可结合肝癌细胞表面的CDH17,并抑制CDH17表达.Lic5 50mg/L组、100mg/L组、小鼠IgG组4 d细胞增殖抑制率在MHCC97H为26.1%、43.6%、6.4%,MHCC97L为26.0%、40.7%、7.7%;Lic5100mg/L组、小鼠IgG组、磷酸盐缓冲液(PBS)组48h细胞迁移抑制率在MHCC97H为36.7%、8.4%、5.6%,MHCC97L为42.3%、10.2%、7.4%(P<0.05);穿膜细胞数在MHCC97H为(39.20±9.56)、(106.50±7.56)、(96.60±13.02)个,MHCC97L为(26.00±8.61)、(86.00±10.26)、(90.40±12.04)(P<0.05);克隆形成数在MHCC97H为(59.30±11.68)、(141.70±19.40)、(150.30±14.64),MHCC97L为(57.20±10.21)、(132.50±9.07)、(121.70±11.93)(P<0.01).Lic5对PLC/PRF/5及MIHA细胞的生物学行为无明显影响.结论 单克隆抗体Lic5能够下调肝癌细胞CDH17表达,抑制肝癌细胞的增殖、迁移、侵袭和克隆形成能力.
目的 觀察抗肝腸鈣粘連蛋白(CDH17)單剋隆抗體Lic5對肝癌細胞生物學行為的影響.方法 Western blot和實時定量聚閤酶鏈反應(PCR)檢測細胞株MHCC97H、MHCC97L、PLC/PRF/5及MIHA中CDH17的錶達.噻唑藍(MTT)比色法、細胞劃痕法、Transwell法及平闆剋隆法檢測Lic5對肝癌細胞生物學行為的影響.結果 CDH17僅在細胞株MHCC97H、MHCC97L中錶達,Lic5可結閤肝癌細胞錶麵的CDH17,併抑製CDH17錶達.Lic5 50mg/L組、100mg/L組、小鼠IgG組4 d細胞增殖抑製率在MHCC97H為26.1%、43.6%、6.4%,MHCC97L為26.0%、40.7%、7.7%;Lic5100mg/L組、小鼠IgG組、燐痠鹽緩遲液(PBS)組48h細胞遷移抑製率在MHCC97H為36.7%、8.4%、5.6%,MHCC97L為42.3%、10.2%、7.4%(P<0.05);穿膜細胞數在MHCC97H為(39.20±9.56)、(106.50±7.56)、(96.60±13.02)箇,MHCC97L為(26.00±8.61)、(86.00±10.26)、(90.40±12.04)(P<0.05);剋隆形成數在MHCC97H為(59.30±11.68)、(141.70±19.40)、(150.30±14.64),MHCC97L為(57.20±10.21)、(132.50±9.07)、(121.70±11.93)(P<0.01).Lic5對PLC/PRF/5及MIHA細胞的生物學行為無明顯影響.結論 單剋隆抗體Lic5能夠下調肝癌細胞CDH17錶達,抑製肝癌細胞的增殖、遷移、侵襲和剋隆形成能力.
목적 관찰항간장개점련단백(CDH17)단극륭항체Lic5대간암세포생물학행위적영향.방법 Western blot화실시정량취합매련반응(PCR)검측세포주MHCC97H、MHCC97L、PLC/PRF/5급MIHA중CDH17적표체.새서람(MTT)비색법、세포화흔법、Transwell법급평판극륭법검측Lic5대간암세포생물학행위적영향.결과 CDH17부재세포주MHCC97H、MHCC97L중표체,Lic5가결합간암세포표면적CDH17,병억제CDH17표체.Lic5 50mg/L조、100mg/L조、소서IgG조4 d세포증식억제솔재MHCC97H위26.1%、43.6%、6.4%,MHCC97L위26.0%、40.7%、7.7%;Lic5100mg/L조、소서IgG조、린산염완충액(PBS)조48h세포천이억제솔재MHCC97H위36.7%、8.4%、5.6%,MHCC97L위42.3%、10.2%、7.4%(P<0.05);천막세포수재MHCC97H위(39.20±9.56)、(106.50±7.56)、(96.60±13.02)개,MHCC97L위(26.00±8.61)、(86.00±10.26)、(90.40±12.04)(P<0.05);극륭형성수재MHCC97H위(59.30±11.68)、(141.70±19.40)、(150.30±14.64),MHCC97L위(57.20±10.21)、(132.50±9.07)、(121.70±11.93)(P<0.01).Lic5대PLC/PRF/5급MIHA세포적생물학행위무명현영향.결론 단극륭항체Lic5능구하조간암세포CDH17표체,억제간암세포적증식、천이、침습화극륭형성능력.
Objective To investigate the effect of monoclonal antibody against liver-intestine cadherin (CDH17) on the cell proliferation, migration, invasion and colony formation of hepatocellular carcinoma cell lines. Methods Hepatocellular carcinoma cell lines MHCC97H, MHCC97L, PLC/PRF/5 and MIHA were examined for CDH17 expression by Western blotting and quantitative real-time polymerase chain reaction (PGR). The combination capacity between bepatocellular carcinoma cell lines and monoclonal antibody Lic5 was detected by the way of immunofluorescence staining. The cell lines were treated with Lic5, PBS and mouse IgG respectively. Methyl thiazol tetrazolium (MTT) assay, wound healing assay, Transwell invasion assay and colony formation assay were used to study the changes in cell proliferation, migration, invasion and colony formation of hepatocellular carcinoma cell lines. Results High expression level of CDH17 was detected in MHCC97H and MHCC97L cell lines. CDH17 protein level was down-regulated but there was no significant effect on CDH17 mRNA after treatment with Lie5 in MHCC97H and MHCC97L. Cellular growth rate of MHCC97H in Lic5 (50 mg/L), Lic5 ( 100 mg/L) and mouse IgG groups was decreased by 26. 1%, 43.6% and 6. 4%, and by 26. 0%, 40. 7% and 7. 7% in MHCC97L on the 4th day respectively (P <0. 05 ). The inhibition rate of cell migration at 48 h was 36. 7%, 8. 4% and 5.6% in Lic5 ( 100 mg/L), mouse IgG and PBS groups in MHCC97H, and 42. 3%, 10. 2% and 7. 4% in MHCC97L respectively ( P < 0. 05 ). The number of invasion cells was ( 39. 20 t 9. 56),(106.50±7.56) and (96.60±13.02) in MHCC97H, and (26.00±8.61), (86.00±10.26) and (90.40±12.04) in MHCC97L in Lic5 (50 mg/L), Lic5 (100 mg/L) and mouse IgG groups, respectively (P < 0. 05 ). The number of colony formation was ( 59. 30 ± 11.68 ), ( 141.70 ± 19. 40 ) and (150.30 ±14.64) in MHCC97H, and (57.20 ± 10.21), (132.50 ±9.07) and (121.70 ±11.93) in MHCC97L in Lie5 (50 mg/L), Lic5 (100 mg/L) and mouse IgG groups, respectively (P< 0. 01 ).There was no significant difference between Lic5 treatment groups and controls in PLC/PRF/5 and MIHA cell lines. Conclusion The anti-CDH17 monoclonal antibody Lic5 can down-regulate CDH17 expression and inhibit the cell proliferation, migration, invasion and colony formation in hepatocellular carcinoma cell lines.